LOCUS V025319 6707 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025319 VERSION V025319 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 941..2197 /label="Sox17(NM_011441)" /note="Sox17(NM_011441)" /gene="Sox17" CDS 2204..2269 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2307..2531 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2577..3005 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3019..3348 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3415..4209 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 4383..4504 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4553..4569) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4577..4593 /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG)." /label="lac operator" promoter complement(4601..4631) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4646..4667 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4955..5540) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5711..6571) /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" promoter complement(6572..6676) /gene="bla" /label="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgagcagcc cggatgcggg 961 atacgccagt gacgaccaga gccagccccg gagcgcgcag cccgcggtga tggcagggtt 1021 gggcccctgt ccctgggccg agtccctgag ccccctcggg gatgtaaagg tgaaaggcga 1081 ggtggtggcg agtagcgggg cgccagccgg gacgtcgggc cgagccaaag cggagtctcg 1141 catccggcgg ccgatgaacg cctttatggt gtgggccaaa gacgaacgca agcggttggc 1201 acagcagaac ccagatctgc acaacgcaga gctaagcaag atgctaggca agtcttggaa 1261 ggcgttgacc ttggcagaga agcggccctt cgtggaagag gccgagcggc tgcgcgtgca 1321 gcatatgcag gaccacccca actacaagta ccggccgcgg cggcgcaagc aggtgaagcg 1381 catgaagcgg gtggagggag gcttcctgca cgctctcgtc gagccccagg ccggcgcgct 1441 tggtcccgag ggcggccgcg tggccatgga tggcctgggt ctgcctttcc cggagccggg 1501 ctatccggcc ggtcctccgc tgatgtctcc gcacatgggc ccccactatc gggactgcca 1561 gggactgggc gctcccgcgc tcgacggcta ccctctgccc actccggaca catccccgct 1621 ggatggcgtg gagcaggacc cggctttctt tgcagccccg ctgccagggg actgcccggc 1681 ggccggcacc tacacttacg ctccagtctc ggactatgca gtgtccgtag agccgcccgc 1741 tggccccatg cgagtggggc cggacccctc gggccctgcg atgccgggga tcctggcgcc 1801 ccccagcgct ctgcacctgt actacggcgc gatgggctcg cccgccgcaa gtgcggggcg 1861 cggtttccac gcgcaacccc agcagccgct gcaaccgcag gcaccgccgc cgccaccgca 1921 gcagcagcac ccagcgcacg gccccgggca accttcgccc cctcccgagg ctctgccctg 1981 ccgggatggc acggaatcca accagcccac tgagctccta ggggaggtgg accgcacgga 2041 attcgaacag tatctgccct ttgtgtataa gcccgagatg ggtcttccct accagggaca 2101 cgactgcgga gtgaacctct cagacagcca cggagccatt tcctccgtgg tgtccgacgc 2161 tagctcagcg gtctactatt gcaactaccc cgacattctc gaggactaca aagaccatga 2221 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc 2281 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg 2341 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata 2401 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt 2461 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt 2521 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc 2581 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac 2641 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt 2701 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc 2761 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc 2821 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact 2881 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg 2941 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc 3001 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca 3061 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca 3121 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc 3181 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc 3241 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta 3301 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga 3361 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt 3421 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat 3481 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag 3541 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac 3601 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac 3661 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc 3721 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg 3781 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag 3841 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat 3901 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag 3961 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc 4021 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg 4081 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg 4141 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag 4201 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat 4261 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc 4321 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc 4381 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca 4441 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat 4501 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc 4561 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 4621 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 4681 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 4741 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 4801 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 4861 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 4921 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 4981 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 5041 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 5101 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 5161 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 5221 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 5281 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 5341 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 5401 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 5461 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc 5521 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 5581 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 5641 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 5701 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 5761 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 5821 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 5881 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 5941 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 6001 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 6061 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 6121 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 6181 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 6241 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 6301 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 6361 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 6421 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 6481 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 6541 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 6601 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 6661 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc //